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Primary culture of fibroblast cells from rat/mouse


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4 replies to this topic

#1 wdyeo

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Posted 11 April 2013 - 12:13 PM

Dear All,
Does anyone have the protocol/paper to culture primary fibroblast cells from the tail or whatever sources of organs/tissues from rat and mouse.
Thank you.

wdyeo

#2 bob1

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Posted 11 April 2013 - 12:26 PM

I don't have one handy but the general procedure is:

Take tissue sample, sterilize surface in bleach (10s) then rinse in sterile water and rinse with 70% ethanol.
Mince tissue into small chunks using sterile scapels (may help to put softer tissues through a cell strainer).
Digest chunks with a protease for some time (tissue dependent).
Neutralize protease and plate out.

#3 Johannes

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Posted 15 April 2013 - 06:12 PM

Bob, just to make sure. Do you mean to sterilize the surface of the tissue sample?

#4 bob1

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Posted 15 April 2013 - 09:07 PM

Sorry, yes, the tissue surface, this is especially important for bits with skin and hair on them, as these are typically covered in bacteria, yeasts and other fungi.

#5 wdyeo

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Posted 16 April 2013 - 03:09 AM

Hi bob1,
Thanks alot for the suggestion...Maybe gotta optimise own protocol :P




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